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. 2023 Mar 9;13:3920. doi: 10.1038/s41598-023-31144-3

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Sfrp2-derived cardiomyocytes form gap junctions. IPSc-derived cardiomyocytes were generated via either the standard broad-spectrum Wnt inhibitor (WntC59) approach or via the Sfrp2 method. After 14 days of differentiation, the cells were analyzed. (a) Cells were delineated by staining for the cell membrane marker wheat germ agglutinin (WGA-white). Cardiomyocytes were visualized with cTnT (green) antibodies and nuclei stained with DAPI (blue). Distribution of gap junctions in cardiomyocytes was determined by immunostaining for connexin-43 (red). N = 3. Scale bar 20 microns. (b) Patch-clamp recordings (Fig. 2e) were analyzed for atrial and ventricular cardiomyocytes. (c) Cells were incubated with antibodies targeting the general cardiomyocyte marker cTNT and the ventricular-specific cardiomyocyte marker MLC2v and then subjected to FACS analysis. The graph shows the number of MLC2v + cells as a percentage of the total cardiomyocyte (cTNT +) population. N = 3. Representative traces are shown.